De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Section: S11-09 Group#: 3 Date Performed: February 7, 2019 Score:

Names: ABORDO, Dirk Karlo LAZARO, Keiana Louise

JAVIER, Jason Alexander SALAZAR, Ruth Angele
SALUD, Nicolas Gabriel TANINGCO, Bea Jane

Laboratory Activity 1
Use of Controls in the Study of Photosynthesis
LABORATORY REPORT EVALUATION FORM
Points Criteria

0 – 10 Introduction:
_________ Clear and appropriate OBJECTIVES and
INTRODUCTION

0-20 Materials and Procedure:

_________ Complete, organized, and images are presented with caption.

Results and Discussion:

Appropriate presentation of DATA and OBSERVATIONS including
chart(s), drawing(s), etc. Accuracy of data.
0-83
_________ Relevant and comprehensive discussion of results. The results are
interpreted correctly. Presents a coherent, well-structured explanation
that accounts for the results or findings of the experiment, making links
between the evidence obtained and existing knowledge. Cites relevant
references correctly. Post lab questions were answered accurately.

Conclusion:
0-5 Clear and concise CONCLUSIONS. Conclusion addressesproblem and
_________ states knowledge gained. Answers to allQUESTIONS.

0-5 Overall – NEATNESS, GRAMMAR, adheres to FORMAT, etc.

_________

______/123 TOTAL POINTS

Teacher Comments:
__________________________________________________________________________________________

Page 1 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

I. Introduction

Being the central biochemical process for every organism, photosynthesis is fundamentally crucial to
sustaining life on earth. Essentially, the word “photo” means “light” and “synthesis” means “to put together”; as
simple as “putting together light” may sound, it involves a highly complex manner that affects a wide-range of
activities. Tracing back to history, it was not until the 1800s that, not only one but many, scientists discovered a
vital natural phenomenon (Photosynthesis Education, n.d.). Starting from a 5-year experiment conducted by Jan
Baptist van Helmont up to Cornelis Van Niel who proposed the general equation for such event is what made
people comprehend the ins and outs of it today. Similar to how humans stack up food in their pantries by going
to a grocery shop, plants, algae, and certain bacteria harness energy from sunlight, carbon dioxide, and water
so that they may churn out food and reinstate oxygen. Only photoautotrophs, those that can utilize energy-
containing molecules from inorganic raw materials, can perform the aforementioned course of action, for they
have a distinctive component in their cells – the chloroplasts. Out of all these organisms, leaves of plants may
be the most ideal to be tested out, for they bestow plenteously amount of surface area for light to be absorbed
due to their broad and flat characteristic; hence, this was the main sample used in this experimentation. Within
this plastid site encloses a pigment called the chlorophyll that is responsible for their green. Although producers
are in the lowest level in the food chain, they influence a wide range of everyday patterns in the global carbon
cycle that includes a lot of complicated and intricate steps. Delving into this further, this complex reaction may
somehow be simplified through this balanced chemical equation:

If, for example, a particular reactant is removed or altered, would the process still be prosperous? How
would the photoautotrophs respond to a certain deficiency? Thereupon, this laboratory activity entitled “Use of
Controls in the Study of Photosynthesis” focused on two types of plants – hydrilla (or hydrilla verticillate) that are
grown underwater and mongo (or vigna radiata) that are grown on land. Even with their different habitats, this
investigation would be of use in upholding the significance of the factors that affect the rate of photosynthesis
(carbon dioxide, water, and sunlight) through examining the variation between one that is grown with exposure
to light and one that is flourished in the dark. Furthermore, two indicators were used for each plant type
respectively: distilled water + bromthymol blue solution that changes its pH level to determine the presence of
carbon dioxide and I2KI solution to detect the existence of starch.
After the experiment, the students have been able to:
1. Pin down and explain the experimental methods in specifying whether or not a specific plant or
leaf has gone through photosynthesis
2. Differentiate the growth rate of plants with respect to the variance of three different factors – light
intensity, carbon dioxide concentration, and temperature
3. Discern the fallout plants underwent once a reactant or an essential component have been
detached from the mechanism
4. Set a point of comparison for further understanding by including a control group and an
experimental group
5. Grasp more firmly the importance of photosynthesis for plants, for people, for the atmosphere,
and for the world in general

Page 2 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

II. Methods and Procedure

A. Initial Preparation

Before the conduction of the experiment, the group intended to prepare materials wherein some were
already readily available in the school’s stock room (marked by an asterisk*) while some were personally brought
by the group members. The materials include the following: (1) 2 250 ml beakers (for the ethyl alcohol and
hydrilla sprigs*, (2) 1000ml beaker (for the distilled water + bromthymol blue solution)*, (3) distilled water +
bromthymol blue solution*, (4) I2KI solution*, (5) wash bottle*, (6) stirring rod*, (7) plastic straw, (8) ethyl alcohol,
(9) 6 test tubes (4 for the carbon dioxide uptake and 2 for the starch production)*, (10) test tube brush*, (11) 4
mongo seedling leaves (4 grown under natural light and 4 in the dark), (12) 2 hydrilla sprigs*, (13) carbon paper*,
(14) wire gauze*, (15) tripod*, (16) bunsen burner*, (17) matchsticks*, (18) 4 petri dishes*, (19) dropper*, (20)
masking tape, (21) marker, and (22) coloring materials.

When the class was instructed to begin, the members prearranged the test tubes through labeling them
with the letters A, B, C, and D for the carbon dioxide uptake experiment and the numbers 1 and 2 for the starch
production experiment using pieces of masking tape and a marker. This is imperative because experiments are
not to be played like a guessing game; the sample should be differentiated from each other easily and accurately
as illustrated in Figure 1.

Figure 1. Labeling the test tubes

Moreover, the group decided to assign different roles to each member so that there would be an
organized flow, and all would be able to take part in the experiment. Specifically, two people were assigned for
each of the two sub procedures and two were held responsible for the documentation, management, and the
recording of observations.

Page 3 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

B. Carbon Dioxide Uptake

In furtherance of the experiment, the class procured an 800ml beaker containing 400ml of a distilled water +
bromthymol blue solution; this is used as an indicator of the presence of dissolved carbon dioxide. Figure 2 below
shows the said solution that all groups of the class used.

Figure 2. Distilled water + bromthymol blue solution

In order to condition the solution in its right color that would be appropriate for determining the
aforementioned gas, two assigned members of the class blew into it using a straw. Also, the teacher reiterated
the caution that it should not be sipped. As carbon dioxide coming from the peoples’ breaths were introduced, a
bubbling phenomenon occurred. Figure 3 displays the said episode.

Figure 3. Blowing into the distilled water + bromthymol blue solution

The members stopped just after the solution turned into an orange-yellow color; such color change
implies that carbon dioxide is already placed within it and is ready to be used for the next step. A member of the
group, then, took Test Tubes A, B, C, and D with him in the designated area. Using a dropper, he managed to
pour equal and identical amounts of such for every test tube, as illustrated in Figure 4, and brought it to the
group’s working area afterward.

Page 4 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Figure 4. Blowing into the distilled water + bromthymol blue solution

Two hydrilla sprigs of the same developmental stage, length, and leaf size were picked from a provided
beaker and placed in Test Tubes A and B one at a time. Test Tubes C and D were out of this conversation thus
containing the distilled water + bromthymol blue solution alone. Figure 5 depicts the time when a member was
situating a hydrilla sprig in Test Tube B, similar to what was done to Test Tube A.

Figure 5. Positioning a hydrilla sprig into Test Tube B

Right after, another group member wrapped around carbon paper into Test Tubes A and C to mimic the
specific incidence of plants being kept or grown in the dark using pieces of masking tape. Figure 6 exhibits the
method as to how this procedure was done.

Figure 6. Wrapping a test tube with carbon paper

Page 5 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

By and by, the members assembled a station wherein three cellular phones, two at the other side and
one at the opposite, with an open flashlight were directly faced to the four test tubes (as seen in Figure 7). While
this “artificial light exposure” set up should have been maintained for a whole hour, the time in the schedule was
running out; hence, it was cut to approximately 37 minutes and 42 seconds.

Figure 7. The arrangement of all the test tubes with exposure to artificial light

Finally, after being able to unwrap the Test Tubes A and C (as shown in Figure 8), the designated data
gatherers began to record the observations. This was done through on-the-spot sketching of the results using
coloring materials and a printed 4-page data sheet.

Figure 8. Unwrapping a test tube covered with carbon paper

Page 6 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

C. Starch Production
The next part of the experiment focused on starch production which began by growing mongo
seedlings under natural light and in the dark right before the day of the experiment took place. Figure 9 shows
where each set was placed; in order for the former to receive a decent amount of sunlight, it was placed
outside while the latter was placed inside a shoebox to prevent any light from coming in. As instructed by the
teacher, they should both be grown at the same time, temperature, soil type, and water conditions.

Figure 9. Growing of mongo seedlings at home

After approximately 5 to 6 days, the cans containing the leaves were brought to school. However, the
plants did not bear enough leaves in such timeframe due to certain conditions, so the group had to ask from
another group who was willing to share. Right after, a member picked fitting 4 mongo seedling leaves from each
type and classified them accordingly as illustrated in Figure 10.

Figure 10. Four mongo seedling leaves from each type

(grown under natural light and in the dark)

Thereafter, a water bath was prepared because ethyl alcohol, having a flammable property, is too critical
to be burned directly. The first step was to pour equal amounts of ethyl alcohol into both Test Tubes 1 and 2.
Figure 11 below displays such act.

Page 7 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Figure 11. Pouring ethyl alcohol into a test tube

The next step was to place the mongo seedling leaves separately in accordance with their type. Test
Tube A contained the leaves that were grown with light and Test Tube B contained the contradictory. Figure 12
manifests a member of the group doing such event.

Figure 12. Positioning the mongo seedling leaves into their respective test tubes
Throughout the time the preceding procedures were being done, the group already arranged a beaker
with 200ml water using the wash bottle, tripod, wire gauze, and bunsen burner, ready to serve as a water bath
for the two test tubes; such is illustrated in Figure 13.

Figure 13. Preparing the primary component of the water bat

Page 8 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

As the water in the beaker had the appropriate temperature of heat to allow the test tubes to be burned,
Test Tubes 1 and 2 were positioned in it as illustrated in Figure 14. This method took around 3-5 minutes long
accompanied by tame shaking in order to distribute the heat.

Figure 14. Test Tubes 1 and 2 in the water bath

After the aforementioned timespan, the heated leaves were moved to the petri dishes as shown in Figure
15. The ethyl alcohol was also drained to a separate petri dish, leaving the leaves alone.

Figure 15. Transferring the samples into separate petri dishes

At last, two drops of I2KI solution, an indicator that changes into a blue-black color in the presence of
starch, was added to the leaves as shown in Figure 16. The challenge for the group was to determine which type
of leaf – the one grown under natural light and in the dark – turned into such color, for it would indicate where
photosynthesis happened. Finally, all the observations were, once again, recorded by the designated members.

Figure 16. Putting I2KI solution into the leaves

Page 9 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

III. Data and Observations

A. Carbon Dioxide Uptake

A.1.
Chemical
Drawing Observations
System

Bromothymol blue, just like what

its name states, is color blue. It
distilled
started as entirely blue and
water + consequently started to show a
bromthymol darker blue shade as water was being
blue added. Likewise, the aforementioned
color dictates that there is no carbon
dioxide present in the pH indicator
yet.

distilled As stated, bromthymol blue is

used to measure the amount of
water + carbon dioxide in a particular
bromthymol environment. When two members of
blue the class blew into the distilled water
(as CO2 is + bromthymol blue (and due to the
blown into fact that the air that we all breathe out
the system) contains carbon dioxide), it changed
in color – from blue to a brown-green.
Moreover, there was an evident
presence of bubbles.

distilled After a while, the distilled water +

bromthymol blue solution changed
water + into an orange-yellow color. From
bromthymol this, it can be inferred that the pH level
blue significantly decreased as a result of
(after CO2 the carbon dioxide being absorbed.
introduction) Among the three chemical systems
above, the state it had in the picture
bears the largest amount of CO2. If
ever that it reverts back to its original
color, then, it may deliver a message
in relation to photosynthesis.

Page 10 of 21
De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

A.2.

Test
Drawing Observations
Tube
Test Tube A contained a hydrilla
sprig and was wrapped with carbon
paper. There was really no change
with the color of the bromthymol blue
after it was exposed to artificial light;
this may be due to some conflicts
within the previous steps. However,
A ideally, since the carbon paper traps
light from being absorbed by the
hydrilla and thus not being able to
conduct photosynthesis, the
bromthymol blue should have reacted
through changing to a clear or light
green color and the leaves should
have turned yellowish.

Test Tube B contained a hydrilla

sprig and was not coated when it was
exposed to artificial light. Thereafter,
there was no color change in the
bromthymol blue because as
mentioned, there was an issue within
B the process. Nonetheless, if ever that
it did go successfully, the group
speculated that the pH indicator would
have turned back to a blue color as a
result of consuming carbon dioxide
and performing photosynthesis.

Test Tube C did not contain any plant,

was covered with carbon paper, and
was subjected to artificial light. As we
know, photosynthesis will only take
place in photoautotrophs like that of
C plant and only with the presence of
chlorophyll. Thereupon, the reason
why there was no color change within
the bromthymol blue is that there was
no particular organism that would use
up the carbon dioxide anyway.

Page 11 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Like Test Tube C, Test Tube D also did

not contain any plant and was put into
artificial light; however, it was not
covered with carbon paper. Although it
was directly getting light, the color of
the bromthymol blue still did not
D
change since there was no plant
involved that could have absorbed the
light and the carbon dioxide.
Generally, photosynthesis will only
take place when a plant, or any kind of
autotroph for that matter, is existent.

B. Starch Production

B.1 Mongo Seedling Leaves Before Heating

Mongo
Seedling Drawing (2 pts each) Observations (2 pts each)
Leaves

Chlorophyll, the pigment that

leaves of absorbs light and turns leaves green, is
Mongo present in these particular leaflings.
When compared side-by-side to such
grown
that were grown in the dark, this appears
under
much darker mainly because of the fact
natural that it was able to capture and adapt to
light process the prolific amount of sunlight at
higher rates.

In order to perform
photosynthesis, plants, most
particularly leaves, must be exposed to
leaves of a satisfactory extent of sunlight with
Mongo respect to the pigment chlorophyll.
grown in Those that were grown in the dark
the dark showed a lighter color of green as
opposed to those that were grown in
the light, for they acclimated
themselves to capture less magnitude
of light.

Page 12 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

B.1 Mongo Seedling Leaves After Heating

Mongo
Seedling Drawing Observations
Leaves
The leaves had a brownish
color feature subsequently after
leaves of heating. It looked like they were slightly
Mongo burned which was actually acceptable
grown since they were heated anyway. The
under group has also noticed that their color
natural got detached and transferred to the
light ethyl alcohol which was once
translucent but now green. The leaves
were also softer than they were before.

One can notice that there is

only a slight difference between the
leaves of leaves that were grown in natural light
Mongo and in the dark. Just like the leaves
grown in that were grown under natural light,
the dark those that were grown in the dark also
had brownish colors and got softer.
Moreover, their original colors faded as
a result of heating and left the green
residue into the ethyl alcohol.

B.3. Mongo Seedling Leaves After Adding I2KI Solution

Mongo
Seedling Drawing (2 pts each) Observations (2 pts each)
Leaves

After adding iodine, the leaves

showed a blue-black color indicating
that there is a presence of starch. In
leaves of
this context, it is imperative to consider
Mongo
that when there is starch,
grown
photosynthesis took place. Granting
under
that these leaves were grown under
natural
the sun, it explicitly showed that they
light
did, in fact, went into the process of
converting sunlight into glucose (and
oxygen).

Page 13 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Although the difference

between the two leaves are not that
distinguishable, there still is a
leaves of distinction. This time, they turned into
Mongo a color of reddish-brown; thus,
grown in implying that they did not go through
the dark an efficient process of photosynthesis.
The reason is pretty obvious – it grew
in the dark and hence did not receive
sunlight, which is a reactant in
producing glucose (and oxygen).

IV. Discussion

A. Carbon Dioxide Uptake

Bromthymol blue is an indicator solution that is commonly used to determine whether or not plants have
expended or released carbon dioxide and underwent photosynthesis. In this experiment, its behavior when air
is blown into it and when there is a presence and an absence of hydrilla sprigs was ascertained. Table 1 shows
the results recorded by the group:

Table 1. Responses of bromthymol blue from blowing

Chemical System Reactions Implication

distilled water + bromthymol blue Blue Absence of carbon dioxide

distilled water + bromthymol blue
(as O2 is blown into the system)
distilled water + bromthymol blue
(after CO2 introduction)
Slow and gradual
Changing into a brown-
presence of carbon
green color & bubbling
dioxide
Changing into an orange-
Carbon dioxide is present
yellow color

It is only with the existence of carbon dioxide that the bromthymol blue would change its color into an
orange-yellow. For that reason, knowing that humans exhale
carbon dioxide, propelling their breaths into it would make that end
reaction possible. Starting from blue, to brown-green, and finally, to
orange-yellow, the chemical system is now equipped to be used as
a determining factor for photosynthesis, for it already holds carbon
dioxide and water (reactants in the aforesaid anabolic chemical
process). By the same token, their changes could be further and
specifically classified through the picture at the left coming from Ms.
Lavenda (n.d.). In this picture, it is important to keep in mind that
the samples were bear unlike the samples in the experiment that
contained distilled water; hence, these are just approximated.
Initially, the bromthymol blue had a pH level somewhere around 7.5
to 8; as two fellow classmates of the members blew into it, it altered
its pH level to somewhere between 6.5 to 7. At last, after all the
Figure 17. Bromothymol Blue pH levels

Page 14 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

carbon dioxide from their breaths (sometimes from the environment too), its pH level was somewhere between
5.5 to 6.

This experiment was also designed to investigate the

response of hydrilla sprigs to artificial light. Although some countries
consider hydrilla as their most problematic plant, for it is responsible
for a number of environmental problems, it is one of the most useful
submerged hydrophytes to use in view of its natural habitat. Due to
the fact that it does not have stomata and that it is grown underwater
(or sometimes in aquariums), looking into how it takes carbon
dioxide as it respires through their whole body surface would be
much easier, for it will make bubbles. To do this, the group prepared
the set up seen in Figure 18.

Figure 18. Test Tubes A, B, C, and D Even though the group was given a choice to use artificial
exposed to artificial light light or sunlight, the cellphones of some of the members were used
since they have the ability to generate light and it would be better
monitored this way. In spite of this, after waiting for around 37 minutes and 42 seconds, the samples did not
show any changes at all; hence, there was an inconvenience in the process – the limited amount of time.
Nevertheless, the group still managed to record the results and made further research, such are organized into
Table 2:

Table 2. Responses of hydrilla sprigs with bromthymol blue & mere bromthymol blue to artificial light
Test
Reaction What should have happened Implication
Tube
The bromthymol blue should have
Remained the Plants cannot perform
A changed to a clear or light green
same photosynthesis without light
color
Remained the The bromthymol blue should have Plants can fortuitously perform
B
same changed to a blue color photosynthesis with light
Remained the For photosynthesis to happen,
C Same yet biased
same there must be plants
Remained the For photosynthesis to happen,
D Same yet biased
same there must be plants

Test Tube A, containing a hydrilla sprig, bromthymol blue, and wrapped with carbon paper, did not have
any substantial reactions after the said interval. Delving into the fact that it was wrapped, jammed of all the light
that should have passed through it, and unexposed to carbon dioxide, the bromthymol blue should have changed
into a clear or light green color. Meanwhile, Test Tube B, containing a hydrilla sprig, bromthymol blue, and
unprotected, should have turned into a blue color that would have indicated that the carbon dioxide has been
taken up. As mentioned in Table 1, when bromthymol blue shows a blue color, there is an absence of carbon
dioxide. It is then safe to say that, carbon dioxide being in the solution and the test tube open to the absorption
of artificial light, the hydrilla sprigs in this sample was well-equipped with all the reactants to undergo
photosynthesis or use up carbon dioxide to produce oxygen. All in all, photosynthesis would only occur if there
is the presence of all the reactants (carbon dioxide & water [within the bromthymol blue] and light).

In addition, Test Tube C, not containing a hydrilla sprig, containing bromthymol blue, and wrapped with
carbon paper, also did not display any changes. The datum in Table 2 says that it is biased even though it did
Page 15 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

have the same outcome with what should have been because from the start, there was an error with the
experimentation. Likewise, Test Tube D, not containing a hydrilla sprig, containing bromthymol blue, and
uncovered, still remained the same. The constant yellow color, then, battles with the belief that only light and
carbon dioxide is needed for photosynthesis to occur. Test Tube C and D proves that photoautotrophs, plants in
particular, should also be present. Simply put, if there are no plants, then there are also no chlorophylls that
would facilitate in the synthesis of carbon dioxide through light. Let’s then say that, for example, all the trees in
the world got cut out, as well as plants, what would happen to our planet? Excluding the other photoautotrophs
like algae being able to photosynthesize, the answer is straightforward — photosynthesis would not happen and
thus oxygen will not be produced in the atmosphere. The world will plunge out into darkness and the animals,
including humans, that temporarily relied on them for sustenance would die (Dove, n.d.).

B. Starch Production

Chiefly, photosynthesis takes place in the green portions of plants or more specifically, in the leaves. The
question is: how do leaves turn green? Well, the pigment called chlorophyll is accountable for all of that with
respect to the precise wavelengths in the visible light spectrum that they reflect. This then brings us to another
question: what really do the leaves color have to do with photosynthesis? After growing the mongo seedlings
(one under natural light and one in the dark) for around 5 to 6 days, their leaves had different shades of green.
Also, the one grown outside looked healthier whilst the other one looked debilitated and similar to a sprouted
mongo (toge). Table 3 shows the results recorded by the group.

Table 3. The colors of mongo seedling leaves

Mongo Seedling Leaves Color Implication

leaves of Mongo grown under natural light Dark green Took up a large amount of sunlight
Took up a small amount of
leaves of Mongo grown in the dark Light green
sunlight

The leaves of the mongo that were grown under natural light resulted to a dark green color while those
that were grown in the dark resulted in a light green color. This shows the relationship between chlorophyll, the
green pigment present in leaves, and its different exposure to light. Chlorophyll just absorbs blue and red light;
it hardly absorbs any green light at all, so the green gets reflected back to our eyes, which is why leaves appear
green (UCSB ScienceLine, 2006). Because of this, it can be alleged that those that were grown in the dark has
less chlorophyll (or none at all) in it thus producing a light green or yellowish color. Correspondingly, it appeared
that the mongo seedlings grown in natural light has loads of chlorophyll and thus, healthier.

In order to fully discern whether or not photosynthesis took

Figure 19. Mongo seedling leaves
in a water bath
place in the said leaves, the first thing that the group did was to put
the mongo seedlings into ethyl alcohol. Through this, the chlorophyll
got extracted and dissolved in order to perceive the results clearly
and hence explaining why the green color of the leaves (both those
grown under natural light and in the dark) got transferred to the
chemical compound which was primarily semi-transparent. The group
also heated the test tubes (see Figure 19) with the purpose of
decolorizing the leaf. Not only that but also by boiling the leaves in
water, it would (1) remove the waxy cuticle that would hinder the
succeeding solution from penetrating, (2) rupture the cell membranes
to make the starch granules in the cytoplasm and chloroplasts
accessible, and (3) convert starch into glucose (Brilliant Biology
Page 16 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

Student, n.d.). Likewise, the second mentioned reason was the one behind the softening of the leaves; the cell
membranes ruptured, and the turgor pressure vanished. This process called “de-starching” should be done in
order to remove the pre-existing starch from the leaves; otherwise, both will show a presence of starch and the
whole experiment would be inaccurate. Table 4 shows the results of such phenomena:

Table 4. The responses of mongo seedlings after heating

Mongo Seedling
Reaction Implication
Leaves

leaves of Mongo Had brown Most of the chlorophyll in the leaves was
grown under natural blemishes, softer, & gotten rid of & the ethyl alcohol turned green
light lost its color from being translucent
Had brown Most of the chlorophyll in the leaves was
leaves of Mongo
blemishes, softer & gotten rid of & the ethyl alcohol turned green
grown in the dark
lost its color from being translucent

The data shows that the mongo seedling leaves grown under natural light and the leaves grown in the
dark did not have discernable differences because both of them looked like they were burned and had lost its
color. If it weren’t for heating, then, the group would have had a hard time in the subsequent step because they
would still have a vibrant color and the reaction of the iodine stain would not be easily seen and discriminated.

The chemical equation of photosynthesis which is 6CO2 + 6H2O + light energy = C6H12O6 + 6O2 can
explain how iodine testing can be useful in demonstrating conclusively the process of photosynthesis in the
mongo seedling leaves samples. In essence, while there is glucose in the product side, some of its excesses
are immediately converted into starch. Specifically, starch granules have been visualized in the stroma of the
chloroplast as well as the cytoplasm (Brilliant Biology Student, n.d.). In a nutshell, it means that when starch is
in there, then, glucose, which is the resulting immediate sugar of photosynthesis, also was. Like the previous
experiment from General Biology 1, the members have learned that with the presence of starch, a sample would
show a blue-black color after putting iodine or I2KI solution for this instance. Table 5 shows the written down
results:

Table 5. The responses of mongo seedlings after adding I2KI solution

Mongo Seedling Leaves Reaction Implication

leaves of Mongo grown Displayed a blue- Contained more starch and executed
under natural light black color photosynthesis proficiently

leaves of Mongo grown in Displayed a reddish- Contained less/no starch and executed
the dark brown color photosynthesis deficiently

After terminating all the chemical processes of the leaves through heating and adding I2KI solution, those
leaves that were grown under natural light turned into a blue-black color and those that were grown in the dark
turned into a reddish-brown color; hence, only the former had adequate starch in it (before was glucose) and
undertook photosynthesis while the latter did not do such efficiently. For that reason, it proves that plants need
to be exposed to sunlight in order to execute the aforesaid chemical process, or for this matter, produce glucose.
Although the mongo seedling in the dark did survive though unexposed, it would not have the capability to
sufficiently excite electrons and use those to create sugars.
Page 17 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

V. Post Laboratory Questions

1. How would you explain the change in the color of distilled water-bromthymol blue solution when
CO2 was introduced in it?

Usually used as a pH indicator, bromthymol blue changes its color once it encounters dissolved carbon
dioxide or other weakly acidic solutions. Because of this characteristic, it is considered as a halochromic material
which makes it an easy determinant of carbon dioxide (Chemicool dictionary, n.d.). Essentially, hence its name,
the indicator started with a blue color or approximately a pH level of 7.5 to 8 (see Figure 1). This means that
carbon dioxide is hardly present in it. Correspondingly, in consonance with prior knowledge, humans exhale
carbon dioxide; thus, in our case, such gas was introduced to the bromthymol blue by means of blowing into the
solution using a straw. Notwithstanding, carbon dioxide does not only react with the bromthymol blue itself but
also with the distilled water, and thereon, forms a weak acid or specifically a carbonic acid (California State
University of Bakersfield, n.d.). During this time, the solution became more acidic, so it shifted into an orange-
yellow color or around a pH level of 5.5 to 6.0 (see Figure 1) indicating that the level of carbon dioxide has
increased. In another light, as the mentioned gas was being intaken, there was a bubbling sensation which
means there is a certain reception and that blowing into it faster would aggravate the reaction further.

2. What will be the resulting color if CO2 is removed from the solution?

As aforesaid, preceding from the blowing procedure, the distilled water-bromthymol blue solution first
started with a blue color which meant that there is imperceptibly any carbon dioxide present in it. Sometime after
blowing, the pH indicator’s condition became more acidic due to carbonic acid, and hence, turned orange-yellow.
Accordingly, when carbon dioxide is withdrawn or is taken up by a certain organism, then, its original color (blue)
would reinstate once again. This kind of operation makes it relatively appropriate for this experiment, for the
students needed a concrete pointer that, for example, the hydrilla sprig really did take up carbon dioxide.
Because such colorless gas is a reactant in photosynthesis, a re-establishment of the solution’s initial color
means that a particular test tube wraps a photosynthetic performance; otherwise, there is a certain factor or
reactant in photosynthesis that was wrongly altered.

3. Why is it necessary to make sure that the sprigs are of the same developmental stage, length
and leaf size?

When conducting an experiment that is directed to compare the same type of plant yet exposed to
different conditions, it is imperative that they are of the same developmental stage, length, and leaf size.
Likewise, this standard of comparison is where the concept of control and experimental set-up emerges.
Categorically, as per the members’ preference, Test Tube B, containing a hydrilla sprig that was not covered by
carbon paper, is in the group because it does not get the variable to be tested — the absence of light. Contrary
to this, Test Tube A, that contained a hydrilla sprig and covered with carbon paper, is included in the experimental
group. However, Test Tube A could still be considered as a control group and Test Tube B as the experimental
group, for the variable to be tested could also be the presence of light. Ultimately, it depends as to how the
experiment performers perceive the activity and how they desire to approach it. To that end, when the two hydrilla
sprigs had a different characteristic, there would be a discrepancy in the collected data since that “distinction”
may have influenced the result; it may not be because of the independent variable alone. Say that the hydrilla
sprig in Test Tube A was two weeks old while the one in Test Tube B was one week old, can one expect that
Test Tube A would instantly turn into a clear or light green color? Also, what if Test Tube A had a smaller leaf
compared to Test Tube B, would that mean the hydrilla sprig in Test Tube B required more carbon dioxide or
not? One could not really know because there is uncertainty and inequality once two things, that are undoubtedly
distinct, are set side by side.

4. Why are the Hydrilla sprigs not placed in the other tubes?

Page 18 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

The fact that photosynthesis works hand in hand with water, carbon dioxide, and sunlight is always brought
up. However, not everyone is well aware of what would happen once one of the most essential yet overlooked
players are removed from the mechanism — plants. In this experiment, Test Tubes C and D did not contain
hydrilla sprigs but were covered with carbon paper and uncovered of such respectively. Therefore, the reason
why hydrilla sprigs were not placed in the other tubes is to prove that there would not be any perceptible changes
in the solution even if all the reactants for photosynthesis is present except for an autotroph itself. Although this
may somehow be obvious, it paves the way for a deeper reason — championing the overall power of
photosynthesis. If there were no plants to undergo photosynthesis, then, all of the carbon dioxides would just
completely retain in the atmosphere and no oxygen would be released. As a result, the whole food chain would
become stationary, the world will completely be deoxidized, and life will not be sustained.

5. Was carbon dioxide utilized by the plant kept in the dark? What experimental observation
indicated this?

Carbon dioxide is a reactant in photosynthesis with the assistance of sunlight; in this case, utilization
means that the carbon dioxide would be used as an ingredient in the aforesaid process. For instance, if there is
insufficient carbon dioxide, a plant cannot photosynthesize, even if it has plenty of light and vice versa. In our
experiment, the mongo seedling that was grown in the dark showed a reddish-brown color which meant that
there is an absence of starch; otherwise, it would have turned into blue-black. As mentioned, this means that
photosynthesis did not happen. Therefore, the said plant did not utilize the carbon dioxide that it engaged with
as it grew. Furthermore, test tube A, the one covered with carbon paper and contained a hydrilla sprig, trapped
all of the artificial light from our cellular phones, a common source of blue light, which potentially could have
initiated photosynthesis. Ideally, it should have turned to a clear or light green color, but there may be something
lacking in the group’s procedure. The group is still unsettled of why the bromthymol blue did not react, but it may
be because some artificial light sources consist wavelengths that are not useful to plants (like green and yellow)
or because the cellular phones were placed improperly. Either way, it did not also utilize carbon dioxide because
there was not any substance to aggravate it to do so. To conclude granting that the plants kept in the dark did
not receive sunlight nor artificial light, an energy provider, at all, the said gas would just solely be absorbed by
the plant – no purpose nor direction.

6. How would you explain the differences in the appearance of the two sets of mongo seedling leaves?

• In the beginning - It can easily be perceived that the one grown under natural light was greener than the
one grown in the dark. As explained, this is because the former’s chlorophyll, the pigment responsible for the
green color of leaves, was absorbing higher rates of sunlight and the latter was deficient of such.
• After heating - There was only a subtle difference between the two mongo seedling leaves that it is almost
imperceptible. It can be then deduced that they are very similar. They both had brown blots, got softer, and
had lost most of their color due to the ethyl alcohol. By the same token, the ethyl alcohol turned light green
from a translucent color thus suggesting that it was where all the leaves’ color got disintegrated.
• After adding I2KI solution - The leaves of the mongo seedlings that were grown in light shifted into a blue-
black color right after such a solution was added to it. This indicates that starch is present in the leaves and
that it went through photosynthesis. In addition, amylose in the starch is responsible for that color (Chemistry
LibreTexts Library, n.d.). On the other hand, the leaves of the mongo seedlings grown without light showed
a reddish-brown color. This implies that the solution just simply stained on the leaves, there is not any
presence of starch, and that most of all, the leaves did not perform photosynthesis.

7. What happens to the ethyl alcohol after heating? Why?

It was clearly demonstrated in the experiment that both variants of leaves (grown under natural light and
in the dark) had disorientation in their color after being heated with ethyl alcohol. Because ethyl alcohol has the
capability to cease chlorophyll from the leaves and circumstantially, kill them. As a result, the ethyl alcohol
somehow inherits the leaves’ color which was again, brought about the chlorophyll pigment that brings about the
Page 19 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

green color. Not only does it become green, but this way, the ethyl alcohol produces carbon dioxide, water, and
heat of around 277.7 kJ/mol as it reacts with oxygen. Ethanol is made of carbon, hydrogen, and oxygen atoms;
hence, the aforementioned outcomes are produced as the molecules uncouple chemical energy stored in the C-
C and C-H bonds. As said by Menihek High School (n.d.), this is considered as an exothermic reaction and has
the chemical equation of:

CH3CH2OH (l) + 3O2 (g) → 3H2O (l) + 2CO2 (g)

ethyl alcohol + 3 molecules of oxygen gas -> 3 molecules of water + 2 molecules of carbon dioxide gas

VI. Conclusion

Science and technology are undoubtedly a rapidly growing field that is shaping the world’s invention and
research. One of the most notable discovery is photosynthesis or the mechanism by which plants, algae and
certain prokaryotes use the energy from sunlight to produce sugar and oxygen essential for all living things.
Accordingly, this experiment helped the students deduce the overall importance of photosynthesis and how to
make sure that all conditions are met for the efficiency of the aforementioned process. To highlight the effect of
the factors involved in the conversion of unusable sunlight energy into usable chemical energy (namely light
intensity, carbon dioxide concentration, and temperature), the leaves or sprigs involved in the experiment were
subjected to solutions that would effectively show various implications.

To determine whether or not the hydrilla sprigs in the first part of the experiment went through
photosynthesis, distilled water-bromthymol blue (or the solution that turns into orange-yellow in the presence of
carbon dioxide) was used. However, some procedures in the experiment were inadequate, so the group had to
think of the possible outcome. They presupposed that the said solution in the test tube containing hydrilla sprigs
but were covered in carbon dioxide (Test Tube A) would turn into a clear or light green color because
photosynthesis could not happen without exposure to artificial light. Meanwhile, the solution in the test tube (Test
Tube B) that was uncovered would regain its blue color; this shows that carbon dioxide has been taken up
through photosynthesis. Delving into this more, the control group and the experimental group may be either from
the two; however, the group deliberated that their control group is within Test Tube B, for the variable which is
the “absence of light” is excluded. Furthermore, the two other test tubes that were bare and did not contain
hydrilla sprigs showed no signs of photosynthesis, for there were no changes in the color of the solution.
However, this is somehow biased because all of the test tubes did not show any changes due to errors.
Specifically, for a more efficient experiment for those who will conduct such in the future, it is recommended that
they leave the said genus of the aquatic plant in direct sunlight as much as possible (or if not, in artificial light)
for a longer amount of time. For better results, the samples should be subjected to light for a whole day. To
conclude, Part A proves that for photosynthesis to happen, plants should receive a sufficient amount of sunlight
and there must be a present plant to absorb the reactants as well.

Besides this, the behavior of mongo seedling leaves that were grown under natural light and in the dark
was also examined. Right after heating them and removing their chlorophyll, I2KI was used to ascertain whether
starch (a remnant of the product glucose) is present. In this setup, the mongo seedling leaves that were kept in
broad daylight displayed a reddish-brown color while one kept in the dark showed a blue-black color; hence, the
earlier showed a presence of starch in its leaves while the latter did not. Thus, an absence of sunlight, no matter
how complete the reactants are, can lead to the inability of any photoautotroph like the mongo seedlings to fully
photosynthesize because sunlight provides the energy for it.

Lastly, this experiment may be used in real-life applications whereas for instance, when planning to
propitiously grow a plant, one must make sure that it is not deprived of any of the following: sunlight, carbon
dioxide, and water. Also, this vivifies the significance of planting trees or preventing deforestation because,
without them, oxygen would not be produced in the atmosphere; as a result, living organisms would die,
structures and the floor would collapse, and the world will experience ultimate catastrophe.

Page 20 of 21
 De La Salle Lipa • Integrated School • Senior High School • Science Learning Area • GENERAL BIOLOGY 2

References:

Brilliant Biology Student. (n.d.). Testing a leaf for the presence of starch. Retrieved from http://brilliantbiolo

gystudent.weebly.com/testing-a-leaf-for-the-presence-of-starch.html

California State University of Bakersfield. (n.d.). Exercise and cellular respirations in the lab. Retrieved from

http://www.csub.edu/chemistry/_files/exerciseandcellularrespirationlabao.pdf

Chemicool dictionary. (n.d.). Definition of halochromism. Retrieved from https://www.chemicool.com/definition/

halochromism.html

Chemistry LibreTexts Library. (n.d.). Starch and Iodine. Retrieved from https://chem.libretexts.org/Bookshelves

/Biological_Chemistry/Supplemental_Modules_(Biological_Chemistry)/Carbohydrates/Case_Studies/

Starch_and_Iodine

Dove, L. L. (n.d.). What if photosynthesis stopped happening? Retrieved from https://science.howstuffworks.co

m/science-vs-myth/what-if/what-if-photosynthesis-stopped.htm

Menihek High School. (n.d.). Hess’s Laws. Retrieved from http://www.menihek.ca/Teacher%20Pages/Teacher

%20Pages/Ruth%20Simmons_files/HesssLaws.pdf

Ms. Lavenda. (n.d.). Bromothymol blue pH tester. Retrieved from http://mslavenda.com/bromothymol_blue.htm

Photosynthesis Education. (n.d.). Discovery of Photosynthesis. Retrieved from https://photosynthesiseducation

.com/discovery-of-photosynthesis/

UCSB ScienceLine. (2006). Why is the chlorophyll in plants green or even red, orange, or brown? Retrieved

from http://scienceline.ucsb.edu/getkey.php?key=1110

Page 21 of 21